AS.Molecular Biology in Hematologic disease.ppt
-
Upload
susi-rutmalem -
Category
Documents
-
view
12 -
download
1
Transcript of AS.Molecular Biology in Hematologic disease.ppt
Agus SuronoOctober 2010
Cytogenetic Genetics Diagnostic Engineering Gene therapy
2Kuliah Biomol 2010
3Kuliah Biomol 2010
Human Genome ProjectsHuman Genome Projects
4Kuliah Biomol 2010
5
Regulation of gene : Regulation of gene :
DNA
mRNA
Nucleus
Cytoplasm
Protein
Intracellular
Extracellular
Membran cell
Transcription
Translation
Kuliah Biomol 2010
DNADNA RNARNA ProteinProtein
transcription translation
Protein SynthesisProtein Synthesis
6
Nucleus Cytoplasm
Kuliah Biomol 2010
7
Protein maturationProtein maturation
Postranslational modification- glycosilation- phosphorylation
Folding
Resite
Kuliah Biomol 2010
8Kuliah Biomol 2010
9
Origin
Copy ????
Kuliah Biomol 2010
10Kuliah Biomol 2010
Autosomal Dominant Autosomal Recessive X-Linked Multifactorial
Identification/DiagnosticIdentification/Diagnostic
11Kuliah Biomol 2010
The haploid human genome has 3 X 109 base pairs of DNA sequence
This contains approximately50,000 - 100,000 genes
(more?)
12Kuliah Biomol 2010
13Kuliah Biomol 2010
14FISH: Fluorescence In Situ Hybridization
Kuliah Biomol 2010
Single Copy DNA (60%) Mainly Euchromatic regions Genes, Pseudogenes
Repetitive DNA (40%) Mainly Heterochromatic regions Tandem repeats
Microsatellite repeats Minisatellite repeats
Interspersed repeats
15Kuliah Biomol 2010
Identify large families with the disease
Clinical Assessment.
Identify a chromosomal locusLinkage.
Refine the candidate intervalRun more markers, construct a contig.
Identify Candidate GenesDatabases, exon trapping, direct selection.
Mutation AnalysisSequencing, southern blotting, SSCP, ....
16Kuliah Biomol 2010
17
INTRODUCTION OF INTRODUCTION OF HEMATOLOGYHEMATOLOGY
Kuliah Biomol 2010
Cellular Components Components of Blood
Red Blood Cells White Blood Cells Inflammatory process Coagulation
Hemorrhage Blood Typing Clotting disorders Transfusion Medicine
18Kuliah Biomol 2010
SpleenSpleen
KidneysKidneys
LiverLiverBoneMarrowBone
Marrow
BloodBlood
HematopoieticSystem
HematopoieticSystem
19Kuliah Biomol 2010
Pluripotent Stem Cell
Myeloid Multipotent Stem Cells
Common LymphoidStem Cells
Unipotent Progenitors
BasophilsEosinophilsNeutrophilsMonocytes
Lymphocytes
ErythrocytesThrombocytes
Erythropoietin
WBC’sRBC’s
Platelets20Kuliah Biomol 2010
Plasma Transport
mechanism 90-92% water. 6-7% proteins 2-3%
Fats Carbohydrates
(glucose) Electrolytes Gases (O2, CO2) Chemical
messengers
Plasma Components
Other3%
Protein7%
Water90%
21Kuliah Biomol 2010
Red Blood Cells Erythrocyte
Hemoglobin – O2 bearing molecule Comprised of 4
subunits: Globin (binds to 1
O2 molecule) Heme (iron)
100% saturation = 4 globin subunits carrying O2
Each gram of hemoglobin = 1.34 ml O2
Cellsalive.com
Components of BloodComponents of Blood
22Kuliah Biomol 2010
Red blood cell production: Erythropoiesis
Erythropoietin
Hemolysis Sequestration Laboratory analysis of
red blood cells Red blood cell count Hematocrit Hemoglobin
Components of BloodComponents of Blood
23Kuliah Biomol 2010
Platelets (Thrombocytes) Megakaryocytes
Thrombopoietin Thrombocytopenia Thrombocytosis
Components of BloodComponents of Blood
24Kuliah Biomol 2010
Class IClass I Class IIClass II Class IIIClass III Class IVClass IV
Blood Loss (ml)Blood Loss (ml) Up to 750Up to 750 750-1500750-1500 1500-20001500-2000 >2000>2000
Blood Loss (% Blood Loss (% volume)volume)
Up to Up to 15%15%
15%-30%15%-30% 30%-40%30%-40% >40%>40%
PulsePulse <100<100 >100>100 >120>120 >140>140
BPBP NormalNormal NormalNormal DecreasedDecreased DecreasedDecreased
Pulse PressurePulse Pressure Normal or Normal or increasedincreased
DecreaseDecreasedd
DecreasedDecreased DecreasedDecreased
Resp. RateResp. Rate 14-2014-20 20-3020-30 30-4030-40 >35>35
Urine Output Urine Output (ml/hr)(ml/hr)
>30>30 20-3020-30 5-155-15 NegligibleNegligible
Mental StatusMental Status Slightly Slightly anxiousanxious
Mildly Mildly anxiousanxious
Anxious, Anxious, confusedconfused
Confused, Confused, lethargiclethargic
Fluid Fluid Replacement Replacement (3:1) rule(3:1) rule
CrystalloiCrystalloidd
CrystalloiCrystalloidd
Crystalloid Crystalloid and bloodand blood
Crystalloid Crystalloid and bloodand blood
25Kuliah Biomol 2010
Blood Types Antigens
A, B, AB, O Rh factor
Rh+ = ~85% Rh- = ~15%
Blood transfusion
26Kuliah Biomol 2010
Blood Typing - ABOBlood Typing - ABO
Blood typeBlood type Antigen Antigen present on present on erythrocyteerythrocyte
Antibody Antibody present in present in
serumserum
OO
ABAB
BB
AA
NoneNone
A and BA and B
BB
AA
Anti-A, Anti-BAnti-A, Anti-B
NoneNone
Anti-AAnti-A
Anti-BAnti-B27Kuliah Biomol 2010
Compatibility Among ABO Blood GroupsCompatibility Among ABO Blood Groups
Reaction with serum of RecipientReaction with serum of RecipientDonor CellsDonor Cells ABAB BB AA OO
ABAB
BB
AA
OO
--
--
--
--
++
--
++
--
++
++
--
--
++
++
++
---= no reaction+ = reaction
Blood Products and Blood Blood Products and Blood Typing Typing
28Kuliah Biomol 2010
Brady; Paramedic Care Principles and Practice
Blood Products and Blood Blood Products and Blood Typing Typing
29Kuliah Biomol 2010
Membrane defect of erythrocytes:
30Kuliah Biomol 2010
31
The Red Cell
Kuliah Biomol 2010
Eritrosit
Hb
Heme + Globin
Fe + Porfirin Chain + non
(,,)
bilirubin
300 juta/sel
SINTESIS Hb & HEMOLISISSINTESIS Hb & HEMOLISIS
Protein (integral, peripheral)
Strength, flexibility (deformability) erythrocytes
Bilayer lipid membrane:
- Protein, lipid, karbohidrat (50,40,10%) - Phospholipid - Attach to cytoskeleton
Membrane defect of erythrocytes:
Protein transmembrane (integral)
- Band 3 (ABO, I, i)
- Glycophorine A (M, N, P. falciparum), B (S, s, U), C- Protein Rh
Function: transport, pump, signal transduction
Protein cytoskeleton (peripheral)
Spectrin , β (band 1,2), actin, ankyrin, band 4.1, band 4.2
Cell stability function: vertikal : spectrin, band 3, ankyrin, protein 4.2 horizontal: spectrin, actin, protein 4.1
Hereditary Spherocytosis
Pathophysiology:
Defect spectrin
Cell membrane is not well constructed
Permeability NaK, dehidration
LIEN‘conditioning effect’
Sferis, fragil
DestructionM, fisik
HEMOLISIS
Membrane defect of erythrocytes:
Band Protei BM /sel Kromosom Gena AA Gena Penyakit
1 Spectri 240240 242242 1q22q231q22q23 80 (Kb)80 (Kb) 24292429 SPTA1SPTA1 HE,HPP,HS,NIHF
2 β Spectri 220220 242242 14q22p24.214q22p24.2 >100>100 21372137 SPTBSPTB HE,HPP,HS,NIHF
2.1 Ankyrin 210210 124124 8p11.28p11.2 >120>120 18801880 ANK1ANK1 HS
2.9 Adduci 103 103 3030 4p16.34p16.3 -- 737737 ADDAADDA -
3* AnExch1 9090 12001200 17q12p2.117q12p2.1 1717 911911 EPB3EPB3 HS,SAO,NIHF
4.1 P4.1 8080 200200 1p33p34.21p33p34.2 >250>250 588588 EL1EL1 HE
4.2 P4.2 7272 250250 15q15q2115q15q21 2020 691691 ELB42ELB42 HS
4.9-8
PAS1 GlypA 3636 -1 rb-1 rb 4q314q31 >40>40 131131 GYPAGYPA
PAS2 GlypC 3232 -200-200 2q144212q14421 1414 128128 GYPCGYPC HE
PAS3 GlypB 2020 -200-200 4q314q31 >30>30 7272 GYPBGYPB
*protein integral; HPP: Hereditary pyropoikilocytosis; NIHF: Nonimmune hydrops fetalis; SAO: Southeast Asian Ovalocytosis
Gene defect protein defect Clinically: hemolytic
Membrane defect of erythrocytes:
Anemia Anemia is a sign, not a separate disease
process. Signs and symptoms may not be present
until the body is stressed. Differentiate chronic anemia from acute
episode. Treat signs and symptoms.
Maximize oxygenation and limit blood loss.
Establish IV therapy if indicated.
38Kuliah Biomol 2010
Brady; Paramedic Care Principles and Practice
Diseases of Diseases of ErythrocytesErythrocytes
39Kuliah Biomol 2010
Sickle Cell Disease Normal red cells maintain
their shape as they pass through the capillaries and release oxygen to the peripheral tissues (upper panel). Hemoglobin polymers form in the sickle cell cells with oxygen release, causing them to deform. The deformed cells block the flow of cells and interrupt the delivery of oxygen to the tissues (lower panel).
Diseases of Diseases of ErythrocytesErythrocytes
40Kuliah Biomol 2010
Sickle Cell Disease (cont.) Sickle cell crises
Vaso-occlusive Musculoskeletal/abdominal pain Priapism Renal/cerebral infarctions
Hematological Lowered hemoglobin Splenic sequestration
Infectious
Management Follow general treatment guidelines prn. Consider analgesics.
Diseases of Diseases of ErythrocytesErythrocytes
41Kuliah Biomol 2010
Polycythemia Overproduction of erythrocytes.
Occurs in patients > 50 years old or with secondary dehydration.
Most deaths due to thrombosis Results in bleeding abnormalities:
Epistaxis, spontaneous bruising, GI bleeding.
Management: Follow general treatment guidelines.
Diseases of Diseases of ErythrocytesErythrocytes
42Kuliah Biomol 2010
THALASSEMIA
HEMOGLOBINOPATHY
Sandoz Atlas: Cinical Hematology, 1988 Gower Medical Publishing 43Kuliah Biomol 2010
Eritrosit
Hb
Heme + globin
Fe + Porfirin Rantai + non (,,)
ThalassemiaThalassemia
300 juta/sel
Sintesis hemoglobinSintesis hemoglobin
2 2 = Hb A >96%2 2 = Hb A2
<3,5%2 2 = Hb F <2%
Molekul Hb (tetramer)
H H eemmee
ThalassemiaThalassemia
Pathophysiology: (Thalassemia )
Defect sintesis of globin
Deformability/liability
Fragilities
LIEN
DestructionM, physic
HEMOLISIS
‘excessive’ of globins Deposited in cell
membrane
ThalassemiaThalassemia
Sp band3 crosslinking
Abnormal Spetcrin association
Abnormal Sp low binding
Deficiency of Spect, Ank Abnormal Sp 4.1 binding
Sp Actin,4.1 instability
Lipid bilayer damage
Excess globin chain Degradat
ion
Denaturation
Band S clustering
Mechanichal removal
Hypercoagulability
Immune removal
“Excessive” of globin result in:
Thalassemia
47Kuliah Biomol 2010
Leukopenia/Neutropenia Too few white blood cells or
neutrophils. Follow general treatment guidelines
and provide supportive care. Leukocytosis
An increase in the number of circulating white blood cells, often due to infection. Leukemoid reaction
48Kuliah Biomol 2010
Leukemia Cancer of hematopoietic cells Initial presentation
Acutely ill, fatigued, febrile and weak, anemic. Thrombocytopenia Often have a secondary infection.
Management Follow general treatment guidelines. Utilize isolation techniques to limit risk of
infection.
49Kuliah Biomol 2010
Lymphomas Cancers of the lymphatic system
Hodgkin's Non-Hodgkins
Presentation Swelling of the lymph nodes Fever, night sweats, anorexia, weight loss,
fatigue, and pruritis
Management Follow general treatment guidelines. Utilize isolation techniques to limit risk of
infection.
50Kuliah Biomol 2010
Multifactorial (viral infection, etc.....)
Complex-genes involvement Chromosomal anomaly :
translocation Diversity on clinical manifestation Therapy approach gene therapy
???Kuliah Biomol 2010 51
Thrombocytosis and Thrombocytopenia Thrombocytosis
An abnormal increase in the number of platelets
Thrombocytopenia An abnormal decrease in the number of
platelets Sequestration Destruction (ITP) Decreased production
Management Provide supportive care and follow general
treatment guidelines.
Clotting DisordersClotting Disorders
52Kuliah Biomol 2010
Hemophilia Deficiency or absence of a blood clotting factor
Deficiency of factor VIII causes hemophilia A. Deficiency of factor IX causes hemophilia B. Deficiency is a sex-linked, inherited disorder.
Defective gene is carried on the X chromosome.
Signs & Symptoms Numerous bruises, deep muscle bleeding, and
joint bleeding.
Clotting DisordersClotting Disorders
53Kuliah Biomol 2010
HEMOPHILIA
Victoria princess family, England 54Kuliah Biomol 2010
• Gene defect/protein FVIII (A), FIX (B)• Bleeding tendency (oozing)• Mild, moderated, severe•Herediter (X-linked) or spontan (approx. 30%)
Role of FVIII/IX in Role of FVIII/IX in hemostasishemostasis
Gene defect FVIII/IX
Deficiency FVIII/IX
Fibrin
Bleeding
Trombosit
Coagulationb c
Hemophilia: defect protein of coagulation factor VIII/IX
Vasokon
str
ksi
A
Hemostasis mechanism
‘close’ the hole
57Kuliah Biomol 2010
58Kuliah Biomol 2010
Gene defect FVIII http://europium.mrc.rpms.ac.uk
1. Gene rearrangement40% hemophilia A severeCross over gene fragmented (meiosis)
2. Single-base substitutions
3. Sequence deletion• >100 kb = severe,
FVIII %, undetected Ag inhibitor?
• <100 kb = mild/moderete
4. Sequence insertion
Management:Causative: factor first! (cryoprecipitate, concentrate, recombinant)
Bleeding correction (surgery)Supportive (transfusion, etc)
Preventive: Marriage counselingDiagnosis of prenatalProphylaxis of bleeding
Complication: Transfusion, inhibitor visibility, high morbidity
Hemophilia (cont.) Management
Treat the patient similarly to others. Administer supplemental oxygen. Establish IV access.
Be alert for recurrent or prolonged bleeding, and prevent additional trauma.
Von Willebrand’s Disease Deficient component of factor VIII
Generally results in excessive bleeding. Generally is not serious; provide supportive
care.
Clotting DisordersClotting Disorders
61Kuliah Biomol 2010
62Kuliah Biomol 2010
Disseminated Intravascular Coagulation System activation of coagulation cascade. Results from sepsis, hypotension, OB
complications, severe tissue or brain injury, cancer, and major hemolytic reactions.
Multiple Myeloma Cancerous disorder of plasma cells. Pathologic fractures are common.
63Kuliah Biomol 2010
64Kuliah Biomol 2010
65Kuliah Biomol 2010
66Kuliah Biomol 2010
67Kuliah Biomol 2010
68Kuliah Biomol 2010
69Kuliah Biomol 2010
70Kuliah Biomol 2010